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A kind of vero cell serum-free culture medium and application thereof

A serum-free medium and cell technology, applied in cell culture active agent, culture process, tissue culture, etc., can solve the problems of microbial contamination, expensive serum, and reduce optimization space

Active Publication Date: 2020-12-01
苏州依科赛生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are many deficiencies in the use of serum: ① there are differences between serum batches, which affect the stability and quality of the vaccine production process; ② the use of serum is easy to cause microbial contamination; Substances that affect cell growth and subsequent downstream purification; ④ Serum is expensive; ⑤ Policies, regulations and industry guidelines are becoming stricter, and the deserumization of biological products is the future trend
At present, most serum-free media developed in China can support cell growth, but most of them add various nutrients, insulin, protein hydrolyzate, etc. on the basis of existing media such as DMEM, DMEM / F12, etc. The quality and safety of the
As disclosed in Chinese patent CN102827804A, it is suitable for the culture medium and method of Vero cell microcarrier suspension amplification culture, the medium includes amino acids, inorganic salts, vitamins, protein hydrolyzate, lipids, buffer components and additives, but the medium is in In the process of culturing Vero cells, it is still necessary to add trace amounts of serum, which cannot be truly serum-free
Moreover, protein hydrolyzate is added to the composition of the medium, the chemical composition is not clear, and there is still potential safety
[0004] However, although the quality of serum-free media from foreign companies such as Life, Thermo, and Lonza is reliable and stable, their price is relatively expensive, there is a risk in the stability of supply, and the ingredients are kept secret, which not only increases the cost of large-scale cell culture, but also reduces the There is room for productivity improvement and cost reduction, and it is not conducive to optimizing and adjusting the medium according to product needs, which greatly reduces the subsequent optimization space

Method used

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  • A kind of vero cell serum-free culture medium and application thereof
  • A kind of vero cell serum-free culture medium and application thereof
  • A kind of vero cell serum-free culture medium and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0291] This embodiment provides a serum-free medium for Vero cell culture, the serum-free medium includes amino acid components, vitamin components, inorganic salt components, trace element components, carbohydrates and other molecular compound components, in,

[0292] The composition of the amino acid component is: L-arginine 45mg / L, L-aspartic acid 30mg / L, L-asparagine 95mg / L, L-cysteine ​​20mg / L, L-glycine 100mg / L L, L-glutamic acid 15mg / L, L-histidine 27mg / L, L-isoleucine 50mg / L, L-leucine 66mg / L, L-lysine 100mg / L, L- Methionine 8mg / L, L-phenylalanine 20m / L, L-proline 15mg / L, L-serine 50mg / L, L-threonine 20mg / L, L-tryptophan 15mg / L, L - Tyrosine 35mg / L and L-valine 30mg / L.

[0293] The composition of vitamin components is: biotin 0.1mg / L, calcium pantothenate 3mg / L, folic acid 10.33mg / L, nicotinamide 15mg / L, vitamin B6 3 mg / L, thiamine hydrochloride 5.5 mg / L, vitamin B12 1.0mg / L, riboflavin 0.9mg / L, choline chloride 35mg / L and inositol 10mg / L.

[0294] The composition ...

Embodiment 2

[0298] This embodiment provides a serum-free medium for Vero cell culture, the serum-free medium includes amino acid components, vitamin components, inorganic salt components, trace element components, carbohydrates and other molecular compound components, in,

[0299] The composition of the amino acid component is: L-arginine 100mg / L, L-aspartic acid 30mg / L, L-asparagine 55mg / L, L-cysteine ​​20mg / L, L-glycine 80mg / L L, L-glutamic acid 15mg / L, L-histidine 27mg / L, L-isoleucine 50mg / L, L-leucine 30mg / L, L-lysine 30mg / L, L- Methionine 30mg / L, L-phenylalanine 30m / L, L-proline 35mg / L, L-serine 50mg / L, L-threonine 20mg / L, L-tryptophan 40mg / L, L - Tyrosine 35mg / L and L-Valine 50mg / L.

[0300] The composition of vitamin components is: biotin 1.0mg / L, calcium pantothenate 8.0mg / L, folic acid 20.33mg / L, nicotinamide 10mg / L, vitamin B6 3 mg / L, thiamine hydrochloride 10 mg / L, vitamin B12 10mg / L, riboflavin 0.9mg / L, choline chloride 35mg / L and inositol 10mg / L.

[0301] The composition ...

Embodiment 3

[0305] This embodiment provides a serum-free medium for Vero cell culture, the serum-free medium includes amino acid components, vitamin components, inorganic salt components, trace element components, carbohydrates and other molecular compound components, in,

[0306] The composition of the amino acid component is: L-arginine 120mg / L, L-aspartic acid 95mg / L, L-asparagine 30mg / L, L-cysteine ​​15mg / L, L-glycine 80mg / L L, L-glutamic acid 15mg / L, L-histidine 27mg / L, L-isoleucine 50mg / L, L-leucine 30mg / L, L-lysine 30mg / L, L- Methionine 30mg / L, L-phenylalanine 30m / L, L-proline 15mg / L, L-serine 15mg / L, L-threonine 20mg / L, L-tryptophan 10mg / L, L - Tyrosine 10mg / L and L-Valine 10mg / L.

[0307] The composition of vitamin components is: biotin 2.0mg / L, calcium pantothenate 5.0mg / L, folic acid 10mg / L, nicotinamide 10mg / L, vitamin B6 10 mg / L, thiamine hydrochloride 5 mg / L, vitamin B12 10mg / L, riboflavin 1.0mg / L, choline chloride 100mg / L and inositol 50mg / L.

[0308] The composition of...

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Abstract

The invention relates to a Vero cell serum-free culture medium and an application thereof. The Vero cell serum-free culture medium comprises an amino acid component, a vitamin component, an inorganicsalt component, trace element components, carbohydrate and other molecular compound components, wherein the trace element components include the following components: zinc sulfate heptahydrate, coppersulfate pentahydrate, ferric nitrate nonahydrate, ferrous sulfate heptahydrate, sodium selenite, nickel chloride, stannous chloride, silver nitrate, cobalt chloride and aluminum chloride. The serum-free culture medium provided by the invention has the advantages of definite chemical components and low cost, avoids the biological safety risk caused by the use of serum when the serum is not needed,and provides convenience for the large-scale production of biological products, especially vaccines. Vero cells grow well in the culture medium, and the serum is not used, so that the stability of the production process of the biological products and the stability of the quality of finished products are improved, and particularly, the research, development and production of novel coronavirus (SARS-CoV-2) vaccines are accelerated.

Description

technical field [0001] The invention belongs to the technical field of cell culture, and in particular relates to a serum-free medium for Vero cells and an application thereof. Background technique [0002] Vero cell (African green kidney cell, Vero cell) is a vaccine production cell line approved by the World Health Organization and my country's biological products regulations. At present, it has been used in the production of various vaccines such as rabies virus and influenza virus. [0003] In vaccine production, in order to achieve high-density culture of cells, 5%-10% serum is usually added to the basal medium. Serum can provide various nutrients required for cell growth, such as hormones, trace elements, adherence factor etc. However, there are many deficiencies in the use of serum: ① there are differences between serum batches, which affect the stability and quality of the vaccine production process; ② the use of serum is easy to cause microbial contamination; Sub...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071
CPCC12N5/0686C12N2500/12C12N2500/14C12N2500/16C12N2500/20C12N2500/22C12N2500/24C12N2500/30C12N2500/32C12N2500/34C12N2500/36C12N2500/38C12N2500/40C12N2500/46C12N2500/90C12N2501/11C12N2501/33
Inventor 徐亮亮陈旭陈刚赵亚王涛董晓惠辛丽丽杨建国
Owner 苏州依科赛生物科技股份有限公司
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